Abstract
Many SARS-CoV-2 neutralizing antibodies (nAbs) lose potency against variants of concern. In this study, we developed 2 strategies to produce mutation-resistant antibodies. First, a yeast library expressing mutant receptor binding domains (RBDs) of the spike protein was utilized to screen for potent nAbs that are least susceptible to viral escape. Among the candidate antibodies, P5-22 displayed ultrahigh potency for virus neutralization as well as an outstanding mutation resistance profile. Additionally, P14-44 and P15-16 were recognized as mutation-resistant antibodies with broad betacoronavirus neutralization properties. P15-16 has only 1 binding hotspot, which is K378 in the RBD of SARS-CoV-2. The crystal structure of the P5-22, P14-44, and RBD ternary complex clarified the unique mechanisms that underlie the excellent mutation resistance profiles of these antibodies. Secondly, polymeric IgG enhanced antibody avidity by eliminating P5-22’s only hotspot, residue F486 in the RBD, thereby potently blocking cell entry by mutant viruses. Structural and functional analyses of antibodies screened using both potency assays and the yeast RBD library revealed rare, ultrapotent, mutation-resistant nAbs against SARS-CoV-2.
Highlights
Neutralizing antibodies, which show efficacy for prophylactic as well as therapeutic use, are being developed against the spike (S) protein of SARS-CoV-2 [1,2,3,4,5,6,7,8]
A yeast library expressing mutant receptor binding domains (RBDs) of the spike protein was utilized to screen for potent neutralizing antibodies (nAbs) that are least susceptible to viral escape
To isolate nAbs, we obtained 44 peripheral blood samples from convalescent patients who had recovered from COVID-19 for approximately 2 months, and isolated peripheral blood mononuclear cells (PBMCs) via Ficoll gradient centrifugation
Summary
Ultrapotent neutralizing antibodies against SARSCoV-2 with a high degree of mutation resistance. Many SARS-CoV-2 neutralizing antibodies (nAbs) lose potency against variants of concern. A yeast library expressing mutant receptor binding domains (RBDs) of the spike protein was utilized to screen for potent nAbs that are least susceptible to viral escape. P5-22 displayed ultrahigh potency for virus neutralization as well as an outstanding mutation resistance profile. P14-44 and P15-16 were recognized as mutation-resistant antibodies with broad betacoronavirus neutralization properties. The crystal structure of the P5-22, P14-44, and RBD ternary complex clarified the unique mechanisms that underlie the excellent mutation resistance profiles of these antibodies. Structural and functional analyses of antibodies screened using both potency assays and the yeast RBD library revealed rare, ultrapotent, mutation-resistant nAbs against SARS-CoV-2
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